# 1437: ALKALIPHILES PIRELLULA MEDIUM (M1PY)


## Main sol. 1437

950	ml	**Solution 1**	
50	ml	**Solution 2**	

1. After autoclaving add solution 2 to solution 1.


## Solution 1

0.25	g	Peptone	
0.25	g	Yeast extract	
5	g	CaCO3	
0.1	g	Na2HPO4 x H2O	
0.5	g	MgSO4 x 7 H2O	
20	ml	**Hutner's salts**	
9	g	Gelrite	
930	ml	Distilled water	

1. Adjust pH to 9.0 with NaOH, autoclave at 121°C for 20 min.


## Solution 2

2	g	N-Acetylglucosamine	
0.2	g	Ampicillin sodium salt	
0.2	g	Cycloheximid	
10	ml	**Vitamin solution**	
40	ml	Distilled water	

1. Adjust pH to 9.0 with NaOH, filter-sterilized


## Vitamin solution

0.2	mg	Vitamin B12	
4	mg	Biotin	
10	mg	Thiamine-HCl x 2 H2O	
10	mg	Calcium pantothenate	
4	mg	Folic acid	
10	mg	Riboflavin	
10	mg	Nicotinamide	
10	mg	p-Aminobenzoic acid	
20	mg	Pyridoxine hydrochloride	
1000	ml	Distilled water	

1. filter-sterilized


## Hutner's salts

10	g	Nitrilotriacetic acid (NTA)	
29.7	g	MgSO4 x 7 H2O	
3.335	g	CaCl2 x 2 H2O	
9.25	mg	(NH4)6Mo7O24 x 4 H2O	
99	mg	FeSO4 x 7 H2O	
50	ml	**"Metals 44"**	
950	ml	Distilled water	

1. Dissolve the nitrilotriacetic acid.
2. Adjust pH to 7.0 with KOH (~7.3 g).
3. Dissolve other salts separately and combine.
4. Adjust pH to 6.8 with NaOH or H2SO4.


## "Metals 44"

250	mg	Na-EDTA	
1095	mg	ZnSO4 x 7 H2O	
500	mg	FeSO4 x 7 H2O	
154	mg	MnSO4 x H2O	
39.2	mg	CuSO4 x 5 H2O	
24.8	mg	Co(NO3)2 x 6 H2O	
17.7	mg	Na2B4O7 x 10 H2O	
1000	ml	Distilled water	

1. Dissolve the EDTA and add a few drops of concentrated H2SO4 to retard precipitation of the heavy metal ions.